The present study was designed to determine the effect of levofloxacin (LVX) treatment on the blood glucose level, insulin sensitivity, anxiety level, nitrite and MDA level of STZ induced diabetic rats. Wistar rats were used in the present study. The rats were made diabetic by the administration of single dose of STZ (45mg/kg, i.p.) and NAD (50mg/kg, i.p.). The rats with the blood glucose level greater than 200mg/dl were considered as diabetic (confirmed at day-3 of STZ-NAD administration). The non-diabetic rats were considered as control and received saline.Diabetic rats received metformin (50mg/kg, p.o.) and LVX (20, 25, 30 and 35mg/kg, i.p.) daily for 14 days (starting from the day at which STZ was injected). Following administration on 14th day,the blood sample was collected and the rats were subjected to behavioral assays for the determination of locomotor activity and anxiety level. Plasma was separated and used for the estimation ofnitrite and malondialdehyde (MDA)level. On 15th day OGTT was performed in the overnight fasted rats for the assessment of insulin sensitivity. The results obtained suggested that the administration of STZ-NAD induced the hyperglycemia at day-3 of administration. Diabetic rats displayed the significant increase in blood glucose, anxiety related behavior, MDA level while significant decrease in the insulin sensitivity and plasma nitrite level. Daily administration of metformin to the diabetic rats decreased the blood glucose level, increased the time spent at the center of open field, reversed the anxiety related behavior in LDT and EPM, did not affect the plasma nitrite level, decreased the plasma MDA level, decreased the fasting glucose level and AUC in OGTT assay. LVX (30 and 35mg/kg) treatment significantly decreased the blood glucose level of diabetic rats. LVX (20, 25 and 30mg/kg) treatment significantly decreased the number of square crossing while LVX (20, 25, 30 and 35) treatment significantly increased the time spent at the center of the field by the diabetic rats. LVX (20 and 35mg/kg) treatment significantly reversed the STZ induced anxiety in LDT while LVX (20, 30 and 35mg/kg) treatment significantly reversed the STZ induced anxiety in EPM test. LVX (20, 25 and 35mg/kg) treatment significantly increased the plasma nitrite level and LVX (20-35mg/kg) treatment significantly decreased the MDA level of diabetic rats. Further only LVX (35mg/kg) treatment significantly decreased the fasting glucose level and increased the AUC of diabetic rats. In conclusion, STZ-NAD administration increased the blood glucose level, anxiety related behavior, decreased the plasma nitrite and increased the MDA level. LVX administration potentiated the diabetogenic effects of STZ-NAD in rats. Daily administration of LVX decreased the blood glucose level of diabetic rats. LVX administration alleviated the STZ induced anxiety in OFT, LDT and EPM test. LVX administration increased the plasma nitrite level and decreased the lipid peroxidation in diabetic rats. The online version contains supplementary material available at 10.1007/s40200-023-01234-0.